【摘要】 目的:研究可溶性组织相容性抗原-G1(sHLA-G1)抑制人自然杀伤细胞(NK细胞)释放穿孔素、颗粒酶,从而降低人NK细胞对猪血管内皮细胞的杀伤作用。方法:利用核转染技术将质粒pcDNA3-sHLA-G1转入LCL721.221细胞株,并提取sHLA-G1。以RT-PCR、斑点酶联免疫吸附(Dot-ELISA)技术分别在基因水平和蛋白水平检测sHLA-G1的表达;以人类NK细胞系(NK92)为效应细胞,猪内皮细胞系PED为靶细胞,用β-己糖氨酶释放实验检测sHLA-G1抑制NK92释放穿孔素、颗粒酶;单四唑(MTT)法检测sHLA-G1抑制NK细胞的杀伤活性。结果:与未加入sHLA-G1的对照组相比,NK92释放的β-己糖氨酶显著减少(P<0.05),且对sHLA-G1组PED的杀伤效率均有明显降低(P<0.01)。结论:sHLA-G1可通过抑制NK92释放穿孔素、颗粒酶以减轻NK92对PED的杀伤作用。更多还原

【Abstract】 Objective: To study the effect of sHLA-G1 inhibiting xenotransplantation rejection through degrading the releasing of porforin and granzyme by NK92. Methods: The recombinant expression vector pcDNA3-sHLAG1 was transfected into the lymphoblastoid cell line LCL721.221 by the nucleofection methods.The expression of sHLA-G1 in the transfected LCL721.221 was detected by RT-PCR and DOT-ELISA technique.NK cell line(NK92) was used as NK effect cells and the porcine aortic endothelial cells line(PED) as targets.The inhibitive effects of the releasing of porforin and granzyme by NK92 was analyzed by the β-hexosaminidase release assay.And the cytotoxicity of NK92 was analyzed by MTT methods. Results:sHLA-G1 conferred a significant degrading the releasing of porforin and granzyme by NK92,and protection PED against NK92 medicated lysis,and the rate of NK92 cell cytotoxicity was reduced to 25.5% in contrast to 71.2% in the control group(P<0.05). Conclusion: sHLA-G1 can degrade the releasing of porforin and granzyme by NK92,and directly protect PED against NK92.These results indicate that sHLA-G1 may be useful to prevent human NK cells to be responsed to porcine xenografts.更多还原